WO2004022735A1 - Procede de production d'une plante a taux de croissance eleve - Google Patents

Procede de production d'une plante a taux de croissance eleve Download PDF

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Publication number
WO2004022735A1
WO2004022735A1 PCT/KR2002/002414 KR0202414W WO2004022735A1 WO 2004022735 A1 WO2004022735 A1 WO 2004022735A1 KR 0202414 W KR0202414 W KR 0202414W WO 2004022735 A1 WO2004022735 A1 WO 2004022735A1
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plant
oshdacl
growth rate
gene
seq
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PCT/KR2002/002414
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English (en)
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In-Cheol Jang
Yoon-Mok Park
Sang-Ik Song
Ju-Kon Kim
Baek-Hie Hahm
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Greengene Biotech Inc.
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Priority to AU2002359024A priority Critical patent/AU2002359024A1/en
Publication of WO2004022735A1 publication Critical patent/WO2004022735A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Definitions

  • the present invention relates to a method for producing a plant having a high growth rate. More specifically, the present invention relates to proteins, OsHDACl ,
  • OsHDAC2 and OsHDAC3 which function as a histone deacetylase, a gene coding for said proteins, and a method for producing a plant having a high growth rate by expressing said gene in the plant.
  • HATs histone acetyltransferases
  • HDACs histone deacetylases
  • the HDACs found in mammals can be classified generally into two classes: the HDACs belonging to class I, composed of approximately 400 to 500 amino acids having a homology with RPD3 as histone deacetylase of yeast, and the HDACs belonging to class II, composed of approximately 1000 amino acids having a homology with yeast's HDA1.
  • HDACs have been very extensively studied and are known only in mammals and yeast, whereas plant HDACs have been known only in that they may be related to the failure of phenotypic function in the Arabidopsis and maize plant.
  • ABA abscisic acid
  • ABA is a plant hormone related to various physiological reactions including those against environmental stresses such as seed generation, dormancy or drought, high concentration of salts and cold. It has been known that abscisic acid plays an important role in maintaining the survival of the plant body by inhibiting the germination and growth of seeds and buds when the plant is under unsuitable growth conditions (see, Seo, M. and Koshiba, T., Trends Plant Sci., 7: 41 -48. 2002). However, in monocot plants including rice, barley, wheat and maize for food production, it is more important that the productivity be increased by. maintaining growth under unsuitable environmental conditions, than the maintenance of survival.
  • OsHDACl among the three kinds of OsHDAC genes isolated from the rice plant, is expressed locally in the whole plant body, and when it is overexpressed, it changes the growth rate and phenotype of the plant and increases the growth of the plant body through increasing its expression even in the presence of ASA which inhibits germination and growth, and thus, completed the present invention.
  • the main purpose of the present invention is to provide protein OsHDACs that function as a histone deacetylase.
  • Another purpose of the present invention is to provide a gene OsHDAC coding for said protein.
  • another purpose of the present invention is to provide a method for producing a plant body having a high growth rate which comprises the step of expressing said OsHDAC genes in the plant body.
  • the present invention relates to a protein, OsHDACl, represented by an amino acid sequence of SEQ. ID. NO. 13 that functions as a histone deacetylase, a gene coding for OsHDACl as defined in SEQ. ID. NO. 13, and an OsHDACl gene coding for OsHDACl as defined in SEQ. ID. NO. 13 and represented by an amino acid sequence of SEQ. ID. NO.l .
  • the present invention also relates to a protein, OsHDAC2, represented by an amino acid sequence of SEQ. ID. NO. 14 that functions as a histone deacetylase, a gene coding for OsHDAC2 as defined in SEQ. ID. NO.
  • the present invention further relates to a protein, OsHDAC3, represented by an amino acid sequence of SEQ. ID. NO. 15 that functions as a histone deacetylase, a gene coding for OsHDAC3 as defined in SEQ. ID. NO. 15, and an OsHDAC3 gene coding for OsHDAC3 as defined in SEQ. ID. NO. 15 and represented by an amino acid sequence of SEQ. ID. NO. 3.
  • the present invention further relates to a method for producing a plant having a high growth rate, which comprises the step of transforming a monocot plant with a recombinant plasmid containing a gene coding for OsHDACl, a gene coding for OsHDAC2, or a gene coding for OsHDAC3 to express the protein OsHDACl, OsHDAC2 or OsHDAC3, respectively.
  • the high growth rate monocot plant produced by method of the present invention is rice, barley, wheat or maize.
  • the method for producing a plant having a high growth rate according the present invention can be further characterized in that the expression of OsHDAC 1 , OsHDAC2 or OsHDAC3 protein is increased by ASA (abscisic acid).
  • the method for producing plant having a high growth rate according to the present invention is also characterized in that the plant exhibits a change in phenotypic properties.
  • the method for producing a plant having a high growth rate according to the present invention comprises the step of transforming a monocot plant with a recombinant plasmid containing a gene as defined in SEQ ID. NO. 1, SEQ. ID. NO. 2 or SEQ. ID. NO.
  • Figure 1 is a drawing which shows the hybridizing position of probes 1, 2, 3, 4 and 5 in OsHDACl, OsHDACl and OsHDACl cDNAs;
  • FIG. 2 is a drawing, which shows the genetic structures of OsHDACl.
  • Figure 3 is a genetic map showing plasmid pAi-OsHDACl
  • Figure 4 is a photograph showing the result of Northern blot analysis, which demonstrates the level of transcription of leaves, roots and callus cells of AB A-treated Ai:: OsHDACl rice plant;
  • Figure 5 is a photograph showing the result of Northern blot analysis, which demonstrates the effect of TSA on the reduction of acetylated H4 in callus cells of Ai:. -OsHDACl rice plant;
  • Figures 6a, 6b, 6c and 6d are photographs showing the growth and morphological change of Ai: . OsHDACl rice plant, the change in the growth of leaves, the change of collars, and the change of roots, respectively;
  • Figures 7a and 7b are the graphs showing the growth of buds and deferential roots of Ai:: OsHDACl rice plant in the absence and presence of ABA, respectively.
  • the present inventors have paid attention to genes or proteins, which control the transcription of genes, particularly HDACs inhibiting the transcription of genes, as the target for changing the growth of plants.
  • genes or proteins which control the transcription of genes, particularly HDACs inhibiting the transcription of genes, as the target for changing the growth of plants.
  • HDACs genes that control the transcription of genes, particularly HDACs inhibiting the transcription of genes.
  • the EST database contains, randomly, the partial base sequences of genes and genomes but does not have any information on the kinds and functions of genes. Therefore, in order to confirm whether the isolated gene actually plays a role or functions as a HDACs, first, a probe was constructed from an isolated gene and then the related genes were separated from a library of rice genes using said probe and amplified.
  • OsHDACl, OsHDACl and OsHDACl genes isolated from EST database are HDACs gene of rice plant. Said three genes have been registered as GenBank accession numbers AF 513382, AF513383 and AF513384, respectively. Then, the present inventors have also observed the growth and phenotypic morphological change of Air.
  • OsHDACl rice which is a rice plant transformed with said OsHDACl. In view of the growth of the rice plant, it was identified that the OsHDACl gene introduced into the transformed Air.
  • OsHDACl rice plant is transcribed at a rate higher than the non-transformed rice plant even under normal conditions, and also is transcribed under the conditions treated with ABA as the growth-inhibiting hormone induced by stresses at a level higher than or comparable to those under normal conditions.
  • ABA the growth-inhibiting hormone induced by stresses at a level higher than or comparable to those under normal conditions.
  • OsHDACl rice plant are much more healthy and strong and provide a higher growth rate, in comparison to those of non-transformed rice plant. From the above results, it could be identified that the plant body having a high growth rate can be developed using the OsHDACl gene.
  • the method for producing the plant body having a high growth rate comprises the step of transforming the monocot plant with a recombinant plasmid containing the OsHDACs gene to express the OsHDACs proteins:
  • the OsHDACs gene is the OsHDACl, OsHDACl or OsHDAC3 gene, and the expression amount of the OsHDACs protein expressed from said gene can be increased by ASA (abscisic acid) as one of the plant hormones, which inhibits germination and growth, to increase the growth rate of the plant body and further to change the phenotypic characteristics.
  • the monocot plants to be transformed according to the present invention can include rice, barley, wheat and maize, etc.
  • the OsHDACs proteins change the structure of chromatin to cause an increase or decrease in the expression of the foreign genes in the genomes, so that the plant body of which the phenotypic characteristics are changed can be produced by controlling the expression amount of the OsHDACs proteins.
  • the OsHDACl gene is expressed locally in the plant and increases the growth rate of the plant by its overexpression and its expression is increased by ABA, this gene can be very efficiently used for producing plants having a high growth rate even under stress conditions including drought, cold, etc., as well as under normal conditions.
  • Example 1 Isolation of genes and construction of probes
  • the sequences in the GenBank database were searched to isolate a few cDNA fragments of about 300 to 400 bp. Then, clones containing said respective cDNA fragments were screened from clones of the EST database of rice and DNAs contained in clones were sequenced by means of base sequence analyzer (Applied Biosystems, USA) to determine finally three kinds of the whole cDNA gene sequences.
  • the sequenced genes were designated as 'OsHDACl (SEQ. ID. NO. 1)', 'OsHDACl (SEQ. ID. NO. 2)' and 'OsHDAC3 (SEQ. ID. NO. 3)', respectively.
  • the probes for searching respective genes in the genome library of the rice plant were constructed.
  • Each of said genetic cDNAs was inserted into Sall-Notl site of pBluescript SKII (Stratagene, USA) to obtain the recombinant plasmid.
  • the recombinant plasmid containing OsHDACl cDNA was cleaved with NcoI/BamHI, Xhol/Notl and Pstl/Notl to obtain DNA fragments of 392 bp (probe 1, SEQ. ID. NO. 4), 1283 bp (probe 2, SEQ. ID. NO. 5), and 415 bp (probe 3, SEQ. ID. NO. 6), as the probes for OsHDACl.
  • the probes for OsHDACl and OsHDAC 3 were obtained using the following primers: ( 1 ) Primers for OsHDACl probes
  • Primer PI 5'-ACGACCCTGACTCTGATATG-3' (SEQ. ID. NO. 7); and Primer Rl : 5'-CCATGGTGTTGGATAATTCT-3' (SEQ. ID. NO. 8) (2) Primers for OsHDAC3 probes
  • Primer P2 5'-CAGCAGCTATGCACCAGAAG-3' (SEQ. ID. NO. 9); and Primer R2: 5'-GCCTCCACGTCCAGTATTGC-3' (SEQ. ID. NO. 10)
  • the product produced from reverse transcription by means of RT-PCR system (Promega, USA) using plasmids containing said primers and OsHDACl or OsHDAC3 was again amplified with PCR to obtain 292 bp (probe 4, SEQ. ID. NO. 1 1) and 249 bp (probe 5, SEQ. ID. NO. 12) DNA fragments, which were respectively separated with electrophoresis and then labeled with [ ⁇ - ,2 P] together with said probe 1, probe 2 and probe 3 for OsHDACl .
  • Figure 1 is the drawing which shows the hybridizing position of probes 1, 2, 3, 4 and 5 in OsHDACl, OsHDACl and OsHDAC3 cDNAs wherein Nc, B, Xh, P, N, S and E denote Ncol, BamHI, Xhol, Pstl, Notl, Sail and EcoRI sites, respectively.
  • the protein sequences of the three kinds of said three cDNA genes were determined.
  • OsHDACl SEQ. ID. NO. 13
  • OsHDAC2 SEQ. ID. NO. 14
  • OsHDAC3 SEQ. ID. NO. 15
  • the proteins are composed respectively of 518, 498 and 510 amino acids and have the molecular weights of 57.5, 55,9 and 56.5 kDa, and preserve 9 domains conserved in mammal HDAC and histidine residue.
  • the separated genes are HDACs gene belonging to class I.
  • Genome database (Beijing Genome Center) of rice plant (Oryza saliva L. ssp. Indica) was screened using OsHDACl cDNA separated above to discover a contiguous sequence 31 135 consistent with 5 '-terminal and 3528-bp of OsHDACl.
  • OsHDACl genomic DNA To determine the remaining portion of OsHDACl genomic DNA, the following primers P3 and R3 were determined from 3-UTR sequence of OsHDACl cDNA using PRIMER DESIGNER 4 program: Primer P3: 5'-GGTGGTGTCTGAATCTCCTA-3' (SEQ. ID. NO. 16); and Primer R3: 5'-AGATGGCATCAGTTACTAAG-3' (SEQ. ID. NO. 17).
  • PCR was conducted using said two primers and genomic DNA of rice plant (Oryza saliva cv. Nipponbare) to amplify genomic DNA of 5.9 kb, and then the amplified DNA was inserted into pGEM-T-Easy vector (Promega, USA) and analyzed for its base sequence.
  • FIG. 2 is a drawing, which shows the genetic structures of OsHDACl, OsHDACl and OsHDAC 3 wherein the black box represents exons. As can be seen from Figure 2, it could be identified that OsHDACl and OsHDAC3 contain 7 exons and 6 introns and OsHDACl contains 6 exons and 5 introns.
  • genomic DNA was extracted from leaves of rice plant (Oryza saliva ca. Nipponbare) grown in greenhouse by means of guanidine-surfactant lysis method using DNAzolES (Molecular Research Center, USA). 5 ⁇ g of genomic DNA was cleaved with 5 kinds of restriction enzymes including Hindlll, EcoRI, Xbal, BamHl and Xhol, and then subjected to electrophoresis on 1.0% (w/v) agarose gel, transferred to hybond N+ nylon membrane (Amersham Pharmacia, USA) and hybridized respectively with probes 1, 2, 3, 4 and 5.
  • restriction enzymes including Hindlll, EcoRI, Xbal, BamHl and Xhol
  • OsHDAC 2 and OsHDAC 3 are the members consisting OsHDACl group and OsHDACl, OsHDACl and OsHDAC 3 are present at a single copy number in rice genome.
  • Example 4 Isolation and Northern blot analysis of RNA In order to investigate the expression patterns of OsHDACl, OsHDACl and
  • RNA was isolated from callus cells, roots and leaves of rice plant using guanidium/LiCl method. 10 ⁇ g of total RNA was denatured in 50% formamide, 2.2 M formaldehyde, 20 mM MOPS [3-(N-morpholino) - propanesulfonic acid] and 0.5 mM EDTA at 70°C for 5 minutes, subjected to electrophoresis on 1 % formaldehyde-agarose gel, transferred to hybond N+ nylon membrane, hybridized respectively with probes 1, 2, 3, 4 and 5, and then analyzed with the same method as in said Southern blot analysis.
  • OsHDAC 2 and OsHDAC 3 are expressed specifically in roots and calluses.
  • Ai promoter as ABA-inducible promoter, composed of ABA (abscisic acid)-reaction complex (ABRC3) from 1.8 kb OsHDACl cDNA and barley HVA22 gene (see, Shen, Q. and Ho, T. H., Plant Cell, 7: 295-307, 1995), CaMV 35S minimal promoter containing only -46 to +100 portion, and 5'-UTR from adhl gene of Arabidopsis was inserted into Clal-Ncol and Ncol-Ncol sites of pSK-RTG promoter (see, Jang, I-C. et al., Mol. Breeding, 5: 453-461, 1999).
  • said plasmid was cleaved with Clal and Notl to obtain 2.2 kb DNA fragment composed of Ai promoter and OsHDACl cDNA, which was then inserted into pSBG-M cleaved with Clal/Notl to construct plasmid (see, Figure 3).
  • Rice plant was transformed with the resulting plasmid using Agrobacterium-mediated transformation and then grown in greenhouse. All of 25 rice plants were produced, and most of them were identified as having a proliferative property and a resistance against 0.5% Vasta solution.
  • Figure 3 is a genetic map showing plasmid pAi-OsHDACl, wherein Ai-P is ABA-inducible promoter; 3 ' pinll is 3'-portion of potato protease inhibitor II gene; 35S is 35S promoter; bar is bar gene to exhibit a herbicide resistance; 3 ' nos is 3 '-portion of nopaline synthase gene; and E, P and N denote EcoRI, Pstl and Notl sites, respectively Southern and Northern blot analysis of the genomes of Ai OsHDACl rice as transduced rice plant was carried out in the same manner as above, except that OsHDACl -specific probe 3 and 586 bp MAR probe b (SEQ. ID. NO.
  • OsHDACl gene is present at 1 to 3 copy numbers in genomic DNA of Ai OsHDACl rice plant and is expressed in all of rice tissues. The subsequent analysis was carried out with selecting the plant having a single copy number of OsHDACl.
  • Example 6 Effect of ABA treatment While leaves of Ai .
  • OsHDACl rice plant produced by above Example 5 was incubated in MS medium with or without 20 ⁇ M ABA, a total RNA was isolated from leaves at regular intervals to examine the expression level of OsHDACl transcription product. As the result, it was identified that the expression of the transcription product reaches at the maximum level one hour after ABA treatment and rapidly decreases after 6 hours. Then, callus cells, leaves and roots of rice plant were incubated in MS medium with or without 20 ⁇ M ABA for one hour, and a total RNA was extracted.
  • Figure 4 is a photograph showing the result of Northern blot analysis, which demonstrates the level of transcription of leaves, roots and callus cells of ABA-treated A i .
  • OsHDACl rice plant wherein NT is untransformed rice plant, and lanes 2, 4 and 6 represent respective individuals of Ai: .
  • OsHDACl rice plant incubated in MS medium with (+) or without (-) 20 ⁇ M ABA for one hour.
  • Figure 4 it could be identified that in case of ABA treatment, the expression level of OsHDACl transcription product in Air.
  • OsHDACl rice plant is 2 to 5 times in leaves, 2 to 6 times in roots, and 9 to 15 times in callus cells, as high as that in untransformed rice plant. Further, the same result could be confirmed from Western blot analysis of OsHDAC l protein.
  • histone was purified from callus (see, Waterborg, J. H. et al., Arch. Biochem. Biophys. 256:167-178, 1987), mixed with the same amount of staining solution (7.4 M urea, 1.4 M NH 3 and 10 mM DTT), and then incubated for 5 hours.
  • the culture was subjected to electrophoresis on acid/urea/Triton (AUT; 1 M acetic acid, 0.5% Triton X-100, 45 mM NH, and 16% acrylamide) comprising the upper layer gel prepared from 1 M acetic acid, 6.3 M urea and 4.4% acrylamide, in a buffer solution containing 0.2 M glycine and 1 M acetic acid.
  • AUT acid/urea/Triton
  • Example 8 Effect of tricostatin A (TSA) treatment
  • Rice callus cells, leaves and roots were incubated in MS medium with (+) or without (-) 20 ⁇ M ABA for one hour, transferred to MS medium containing 20 ⁇ M ABA and 1 ⁇ M TSA, incubated for 1 to 6 hours and then subjected to TSA treatment.
  • rice tissues was pulverized, homogenized with a buffer solution containing 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 10% glycerol, 0.5% Triton X-100, 2 mM phenylmethanesulfonyl fluoride, 1 ⁇ g/ml aprotinine, 1 ⁇ g/ml pepstatin and 1 ⁇ g/ml leupeptin, and then incubated at 4°C for one hour. After incubation, the mixed solution was centrifuged at 4°C for 5 minutes, and the protein concentration was determined using Bradford solution (BioRad, USA).
  • the protein extract was separated from 12%) SDS gel, transferred to polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore), and then reacted with mouse HDAC1 (mHDACl), histone H4 (H4), acetylated histone (AcH4), the secondary antibody wherein primary antibody to acetylated histone H4 (AcH4) (Upstate Biotechnology, USA) is combined with alkaline phosphatase.
  • PVDF polyvinylidene difluoride
  • Figure 5 is a photograph showing the result of Northern blot analysis, which demonstrates the effect of TSA on the reduction of acetylated H4 in callus cells of Ai:: OsHDACl rice plant in which NT is callus cells of untransformed rice plant, AcH4 is acetylated histone H4, H4 is total histone H4, and PC is a core histone separated from chicken red cells (Upstate Biotechnology, USA).
  • NT callus cells of untransformed rice plant
  • AcH4 is acetylated histone H4
  • H4 is total histone H4
  • PC is a core histone separated from chicken red cells (Upstate Biotechnology, USA).
  • FIG 5 it could be identified that the expression level of tetra-acetylated histone H4 is greatly increased in both of Ai: . OsHDACl rice plant and untransformed rice plant after TSA treatment but total H4 is substantially not changed.
  • OsHDACl rice plant Air.
  • OsHDACl rice plant was incubated as the first generation and then, young seedlings or calluses at the second generation were transferred to MS medium with or without 20 ⁇ M ABA and incubated for 2 weeks. After incubation, the whole shape of rice plant, and leaves, collars and roots were observed. As the control group, untransformed rice plant was also observed.
  • Figures 6a, 6b, 6c and 6d are photographs showing the growth and morphological change of Air. OsHDACl rice plant, the change in the growth of leaves, the change of collars and the change of roots, respectively.
  • NT denotes untransformed rice plant and lanes 2, 3 and 4 in Figures 6b, 6c and 6d denote respective individuals of transformed Air.
  • OsHDACl rice plant As can be seen from Figure 6a, untransformed plant exhibited the inhibition of growth in the presence of ABA and was killed after 2 to 3 weeks, whereas Air. OsHDACl rice plant exhibits an increase of growth in the presence of ABA. Further, as can be seen from Figures 6b, 6c and 6d, leaves, collars and roots of Air. OsHDACl were very healthy and strong in comparison to those of untransformed rice plant.
  • FIGS 7a and 7b are the graphs comparatively showing the growths of buds and deferential roots of Air.
  • the seeds of Ai: when the plant was incubated in MS medium containing 20 ⁇ M ABA, the seeds of Ai: .
  • OsHDACl rice plant exhibited a very rapid rate of bud germination and deferential root growth in comparison to that of the seeds of untransformed rice.
  • the rice plant transformed with the control vector not containing OsHDACl gene did not show the characteristics shown in Ai: OsHDACl rice plant.
  • the present invention relates to a protein having a function of histone deacetylase, i.e. OsHDACl, OsHDAC2 and OsHDAC3, a gene coding for said proteins, and a method for producing a plant having a high growth rate by expressing said gene in the plant.
  • OsHDACs proteins change the structure of chromatin to increase or decrease the expression of a foreign gene in genomes, so that the expression amount of OsHDACs proteins can be controlled to produce the plant having varied phenotypic characteristics.
  • OsHDACl gene is expressed locally in the plant and increases the growth rate of plant by its overexpression and its expression is increased by ABA. Therefore, it can be very efficiently used for producing plants having a high growth rate even under stress conditions including drought, cold, etc., as well as under the normal conditions.

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Abstract

La présente invention concerne des protéines OsHDAC1, OsHDAC2 et OsHDAC3 à activité histone-désacétylase, un gène codant pour ces protéines, ainsi qu'un procédé de production d'une plante à taux de croissance élevé par expression dudit gène dans la plante. Selon l'invention, les protéines OsHDAC modifient la structure de la chromatine pour augmenter ou réduire l'expression d'un gène étranger dans les génomes, de sorte que la quantité d'expression des protéines OsHDAC peut être régulée pour produire une plante possédant des caractéristiques phénotypiques variées. En particulier, la protéine OsHDAC1 est exprimée localement dans la plante et augmente le taux de croissance de celle-ci par sa surexpression et son expression est accrue par ABA. De ce fait, cette protéine peut être utilisée de façon particulièrement efficace pour produire des plantes à taux de croissance élevé, y compris dans des conditions difficiles, notamment de sécheresse, de froid, etc., ou dans des conditions normales.
PCT/KR2002/002414 2002-09-05 2002-12-23 Procede de production d'une plante a taux de croissance eleve WO2004022735A1 (fr)

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KR10-2002-0053637A KR100510211B1 (ko) 2002-09-05 2002-09-05 높은 성장율을 갖는 식물체의 생산방법
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Publication number Priority date Publication date Assignee Title
WO2014118123A1 (fr) 2013-01-29 2014-08-07 The University Court Of The University Of Glasgow Procédés et moyens pour augmenter la tolérance aux contraintes et la biomasse dans des plantes
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WO2017009253A1 (fr) * 2015-07-10 2017-01-19 The University Court Of The University Of Glasgow Procédés et moyens pour augmenter la tolérance au stress et la biomasse dans des plantes
US11350614B2 (en) 2017-06-19 2022-06-07 Biocytogen Pharmaceuticals (Beijing) Co., Ltd. Genetically modified non-human animal with human or chimeric CD28

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CN112409466B (zh) * 2019-08-21 2023-05-09 中国科学院微生物研究所 蛋白质hda703在调控水稻产量中的应用

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Publication number Priority date Publication date Assignee Title
WO2014118123A1 (fr) 2013-01-29 2014-08-07 The University Court Of The University Of Glasgow Procédés et moyens pour augmenter la tolérance aux contraintes et la biomasse dans des plantes
CN104774826A (zh) * 2015-03-20 2015-07-15 中国科学院华南植物园 一种组蛋白脱乙酰化酶及其编码基因和应用
CN104774826B (zh) * 2015-03-20 2017-12-12 中国科学院华南植物园 一种组蛋白脱乙酰化酶及其编码基因和应用
WO2017009253A1 (fr) * 2015-07-10 2017-01-19 The University Court Of The University Of Glasgow Procédés et moyens pour augmenter la tolérance au stress et la biomasse dans des plantes
US11350614B2 (en) 2017-06-19 2022-06-07 Biocytogen Pharmaceuticals (Beijing) Co., Ltd. Genetically modified non-human animal with human or chimeric CD28

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US7060873B2 (en) 2006-06-13

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